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Physics Faculty Publications Physics
12-13-2004
Medical image processing using transient Fourier
holography in bacteriorhodopsin flms
Sri-Rajasekhar Kothapalli
University of Massachusets Boston
Pengfei Wu
University of Massachusets Boston
Chandra S. Yelleswarapu
University of Massachusets Boston, chandra.yelleswarapu@umb.edu
D.V.G.L.N. Rao
University of Massachusets Boston, raod@umb.edu
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Recommended Citation
Kothapalli, Sri-Rajasekhar; Wu, Pengfei; Yelleswarapu, Chandra S.; and Rao, D.V.G.L.N., "Medical image processing using transient
Fourier holography in bacteriorhodopsin flms" (2004). Physics Faculty Publications. Paper 3.
htp://scholarworks.umb.edu/physics_faculty_pubs/3
Medical image processing using transient Fourier holography in
bacteriorhodopsin films
Sri-Rajasekhar Kothapalli, Pengfei Wu, Chandra S. Yelleswarapu, and D. V. G. L. N. Rao

Citation: Appl. Phys. Lett. 85, 5836 (2004); doi: 10.1063/1.1833567
View online: http://dx.doi.org/10.1063/1.1833567
View Table of Contents: http://apl.aip.org/resource/1/APPLAB/v85/i24
Published by the American Institute of Physics.

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Medical image processing using transient Fourier holography
in bacteriorhodopsin lms
Sri-Rajasekhar Kothapalli, Pengfei Wu, Chandra S. Yelleswarapu, and D. V. G. L. N. Rao
a)
Department of Physics, University of Massachusetts, Boston, Massachusetts, 02125
(Received 19 March 2004; accepted 16 October 2004)
Real time image processing is demonstrated by recording and reconstructing the transient
photoisomerizative grating formed in the bR lm using Fourier holography. Desired spatial
frequencies including both high and low band in the object beam are reconstructed by controlling
the reference beam intensity. The results are in agreement with a theoretical model based on
photoisomerization grating. We exploit this technique to process mammograms in real-time for
identication of microcalcications buried in the soft tissue for early detection of breast cancer. A
feature of the technique is the ability to transient display of selected spatial frequencies in the
reconstructing process which enables the radiologists to study the features of interest. 2004
American Institute of Physics. [DOI: 10.1063/1.1833567]
Real-time optical information processing using nonlinear
optical materials has received considerable attention recently
due to ever increasing demand for processing a huge amount
of data.
13
Photorefractive materials are widely used for
implementation of optical Fourier transform operations such
as edge enhancement, band pass ltering, noise removal, and
pattern recognition.
4,5
There is much scientic interest in
photoisomerizative biological and organic molecules for
studies on optical nonlinearity
6
and applications in photonics
such as optical storage,
1
switching,
7
and information
processing.
8
Small molecular size with resulting high resolu-
tion and low driving energy make them an ideal choice for
applications in optical image processing as compared to the
inorganic photorefractive materials. The biological photo-
chrome bacteriorhodopsin shows many intrinsic optical and
physical properties
9
and is receiving much attention as an
alternative to conventional photorefractive materials for ap-
plications in optical recording and image processing. The
spatial ltering in bR was performed using a control beam
that precisely manipulates spatial frequencies exploiting
saturable absorption
10
and photoinduced aniosotropy.
8
As
compared to other materials like photorefractive crystals, the
bR lms have many advantages such as high spatial resolu-
tion of over 6000 lines/mm,
11
high photoisomerization ef-
ciency, ease of preparation of thin lms of large area, and
reversibility of use.
In this letter, we study transient Fourier holographic
gratings based on photoinduced isomerization properties of
bR lms. The desired spatial frequency components in the
Fourier transformation of the object can be selected by
matching their intensity to that of the reference beam. We
used this technique to process mammograms in real-time for
identication of microcalcications buried in the soft tissue.
The results offer useful information to radiologists for early
detection of breast cancer. An added advantage is that the
radiologist can compare the processed image and original
image in real-time by blocking and unblocking the object
beam.
Figure 1 shows the experimental arrangement of Fourier
holography for study of transient nonlinear spatial ltering
and applications of image processing. The 568 nm output
from an ArKr ion laser is expanded by a beam expander
(BE), and then split into two beams by a 5050 beam splitter
BS
1
. The object beam passing through an object (mammo-
gram or binary object E), is transformed to Fourier plane by
a lens L
1
with focal length of 20 cm. At the Fourier plane the
bR lm is placed for real-time processing of spatial fre-
quency information contained in the object. The Fourier
spectrum of the object contains low spatial frequencies at the
center with high intensity and high spatial frequencies at the
edge with low intensities. The bR lm, wild-type purchased
from Munich Innovative Biomaterials GmbH, has optical
density of about 5 at 570 nm with a thickness of about 100
m. The reference beam overlaps the Fourier transform of
the object beam on the bR lm using beam splitter BS
2
thereby recording a Fourier hologram. A small angle of about
5 between reference and object beams is set to give maxi-
mum diffraction efciency. A variable attenuator (VA) is
placed in the reference path to match its intensity to that of
the desired spatial frequency band in Fourier spectrum of the
object beam. It takes about 5 s to saturate the grating with
optimum modulation and the hologram is completely re-
corded. When the object beam is blocked, for the readout
process, the reference beam performs the reconstruction of
the recorded Fourier hologram. During the readout process
the reference beam erases (in about 10 s) the contrast in the
population grating and thus bR lm is brought back to its
original state with no history of object information and is
ready for the next cycle. The desired frequency band of the
object wave reconstructed from the hologram passes through
the lens L
2
with an inverse Fourier transform conguration
and is imaged on to the CCD camera.
Initially we investigated temporal and intensity-
dependent properties of holographic grating recorded by us-
a)
Electronic mail: raod@umb.edu
FIG. 1. Experimental arrangement for study of edge enhancement using
transient Fourier hologram recorded in bR lm. M: mirrors; BS: beam split-
ters; and L: lenses.
APPLIED PHYSICS LETTERS VOLUME 85, NUMBER 24 13 DECEMBER 2004
0003-6951/2004/85(24)/5836/3/$22.00 2004 American Institute of Physics 5836
Downloaded 29 Mar 2012 to 158.121.194.143. Redistribution subject to AIP license or copyright; see http://apl.aip.org/about/rights_and_permissions
ing bR lms. For this study we slightly modied the experi-
mental setup shown in Fig. 1. The object and Fourier
transform lenses are removed from the object path and the
CCD camera is replaced by a photodetector. The intensity in
the reference beam is xed at 26 mW/ cm
2
while intensity in
the object beam is varied from 1.6 mW/ cm
2
to 1.6 W/ cm
2
,
to stimulate the range of intensities of spatial frequencies in
the Fourier spectrum of the object. The holographic grating
formed due to the interference between the two beams is
recorded in bR lm for about 5 s to reach its saturation. The
optimum of diffraction efciency occurs when object beam
intensity is matched to the reference beam intensity. When
the object beam is blocked, the reference beam decreases the
grating contrast decreases due to isomerization process from
M to B resulting in decay of the diffracted signal. Figure 2
shows the decay of diffraction signal with time for an object
beam intensity of 26 mW/ cm
2
and matched reference beam
intensity of same value. We observed decay of the diffraction
signal for a series of object beam intensities. The values of
diffraction signals obtained from this data are plotted as a
function of object beam intensities for various time delays
shown in Fig. 2 inlet. It is clear that the diffraction efcien-
cies show optimum values when the object beam intensity
matches the reference beam. At either low or high intensity
region of the object beam, the diffraction efciency de-
creases.
The experimental results can be understood using the
mechanism of photoinduced isomerization grating formed in
bR lm. Upon excitation of the initial B state (all trans), the
bR molecule goes through several intermediate states with
short lifetimes to the relatively long-lived M state (cis) and
relaxes back to the initial state spontaneously.
12
We may con-
sider only the B and M states as the other intermediate states
in the photocycle are short lived. The initial B state has an
absorption peak at 570 nm while the long-lived M state has
an absorption peak at 412 nm.
Monochromatic plane waves are used to form holo-
graphic grating in bR lm. Thus, the bR molecules simply
undergo photoisomerization process as follows:
B
R
2
,R
T
R
1
M,
where R
i
is the rate of the photoisomerization, i =1 and 2
corresponding to the photoisomerizations from BM (trans
to cis) and MB (cis to trans) respectively while R
T
is the
rate of the thermal isomerization from MB. Due to the
spatial intensity modulation induced by the interference of
the two recording beams in the sample, the photoisomeriza-
tion from BM is spatially modulated leading to a M state
population grating. The diffraction efciency of this grating
is proportional to the square of modulated contrast of the M
state population grating. Following the theoretical model for
biphoton grating in azo materials,
13
we express the temporal
behavior of the modulated contrast in the reading process as
N
2
t =
A
1
A
2
IN
0
1 + A
1
+ A
2
I
2
e
1+A
2
IR
T
t
, 1
where N
0
is the total number of bR molecules given by N
0
=N
1
t +N
2
t, N
1
t and N
2
t being the temporal popula-
tions of the M and B, respectively. R
i
/ R
T
=A
i
I, where A
i
=
i

i
/ qR
T
,
i
is the absorption cross section of the B state
i =1 or M-state i =2,
i
is the quantum yield of photoi-
somerization from BM state i =1 and MB i =2 and
is the angular frequency of the light. From the Eq. (1) it
follows that the dependence of both the grating contrast and
decay time of diffraction signal on reference beam intensity I
is in agreement with our experimental observation. In addi-
tion, our experimental results of the diffraction decay can be
explained in terms of two lifetimes. For example, theoretical
plot (solid line) in Fig. 2 is obtained using two lifetimes, 62
and 240 ms. We believe the short one originates from the
isomerization rate from MB, and the long one is due to the
mechanism of bR molecular reorientation. We modied Eq.
(1) with two exponential decays corresponding to these two
lifetimes to t the experimental data in Fig. 2. Diffraction
decay data for other object intensities can also be explained
similarly.
The intensity in Fourier spectrum of any object is spa-
tially distributedlow frequencies are at high intensity and
high frequencies are at low intensity. We exploit the temporal
and intensity dependent features of diffraction efciency of
bR sample for selective recording of spatial frequencies for
applications in image processing. To illustrate the feasibility
of the technique we rst recorded high spatial frequency
components of a binary object E, by matching the corre-
sponding intensity to the intensity of the reference beam. The
recording process takes about 5 s so that the contrast of Fou-
rier hologram reaches its maximum in the bR lm. When the
object beam is blocked, the reconstructed wave by the refer-
ence beam shows an edge enhancement effect. The experi-
mental results are shown in Fig. 3(b). In addition, the bR
material also has saturable absorption feature which enables
it to directly pass low-frequency components of the image
and block high-frequency components. In the absence of ref-
erence beam, an image with soft edge of object E as shown
in Fig. 3(c) is obtained. Thus this scheme is able to give the
processed results of both low-frequency and high-frequency
components in real time and can be adopted for other fre-
quency bands as well.
We now exploit this scheme for processing mammo-
grams for detection of microcalcications. The microcalci-
cations (tiny calcium deposits in human breast) correspond
to high spatial frequencies in the Fourier spectrum because
of their small size 10 m and diffuse nature. The object
E in the above experiment is replaced with region of interest
(ROI marked by the radiologist) of the mammogram as de-
picted in Fig. 3(d). By controlling the reference intensity,
edge enhancement of mammograms is achieved leading to
FIG. 2. Experimental results of temporal decay of diffraction signal at
matched object and reference beam intensity of 26 mW/ cm
2
. The solid line
is the theoretical t. Inlet: diffraction signal as a function of object beam
intensity for various time delays with xed reference beam intensity of
26 mW/ cm
2
.
Appl. Phys. Lett., Vol. 85, No. 24, 13 December 2004 Kothapalli et al. 5837
Downloaded 29 Mar 2012 to 158.121.194.143. Redistribution subject to AIP license or copyright; see http://apl.aip.org/about/rights_and_permissions
clear display of microcalcications (not visible to the naked
eye of the radiologist) on the monitor as shown in Fig. 3(f).
Irrespective of different size of regions of interest and den-
sity of soft tissue present in mammograms, the radiologist
can easily scan for desired microcalcication clusters by ro-
tating the variable attenuator placed in the path of the refer-
ence beam. With the spatial resolution of bR lm
6000 lp/ mm
11
and CCD camera resolution
67 lp/ mm we could easily detect microcalcications of
10 m in size (1012 lp/mm).
An additional attractive feature of this technique is the
transient nature in display of different spatial frequencies in
the Fourier spectrum of an object. Abnormalities detected in
mammography are classied as spiculated masses, stellate
lesions, circumscribed masses, and microcalcications. As
these abnormalities vary in size and intensity they corre-
spond to different spatial frequency bands in the Fourier
spectrum of the mammogram. From the inlet in Fig. 2 we
can infer that at a given time, all the spatial frequencies exist
with different diffraction efciencies. The maximum ef-
ciency occurs for a selected band of frequencies that are
optimized by matching the intensities of the object and ref-
erence beams. Other spatial frequencies also appear but are
lower intensity. The dominant frequency band persists for a
relatively longer time whereas the other frequencies decay
fast. Thus we can distinguish between different spatial fre-
quencies as they reveal at different times. We recorded a
movie of the whole hologram recording and readout process
using a fast CCD so that the readout process can revisit as
well as freeze the frames for the radiologist to concentrate on
a particular band of frequencies. To demonstrate this concept
we used a resolution chart (USAF negative target, Edmund
Optics). As shown in Fig. 4(a) at time t =0 we can observe all
the frequency groups [(A) low frequency group, (B) middle
frequency group, and (C) high frequency group] but at time
at t =5 only the frequency group B which matches the refer-
ence beam intensity remains clear while other frequency
groups vanish. The radiologist also has the choice of select-
ing the dominant frequency band by varying the reference
beam intensity during the whole process. This could be a
potential advantage to the radiologist in diagnostics of abnor-
malities in the mammogram that occur at different scales
such as clusters of microcalcications at small scales, edges
of smooth or star-shaped objects in the breast at middle
scales and architectural distortions at large scales.
14
In conclusion, interesting results on temporal and
intensity-dependent properties of Fourier holographic grat-
ings using bR lms are reported. The analysis using a theo-
retical model based on photoinduced isomerization agrees
with our experimental results. We exploit this technique to
process mammograms in real-time for detection of microcal-
cications buried in the background of soft tissue. The re-
sults are useful to radiologists for early diagnostics of breast
cancer. In addition, we found that the information in static
processed images can be further separated in time scales.
These temporal image features offer useful information to
the radiologist about the pathological changes in the image
which cannot be obtained from the static processed image.
This optical technique can be easily adopted for processing
digital mammograms by replacing the object in Fig. 1 with
SLM connected to the computer.
This work is supported by National Cancer Institute,
NIH, Grant No. 1R21CA89673-01A1. The high quality pho-
tographic images of the clinical mammograms were obtained
from the University of Massachusetts Medical School,
Worcester, MA. We thank Dr. Carl DOrsi and Professor An-
drew Karellas for providing them. We are grateful to Dr.
Joby Joseph, I. I. T. Delhi, India, for valuable discussions.
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FIG. 3. Experimental results of image processing using Fourier holography.
For the binary image E: (a) original, (b) edge-enhanced, and (c) edge-
softened. For the mammograms: (d) original image with of region of interest
(ROI) circled by radiologists, (e) blowup of the ROI; (f) processed image
showing calcications.
FIG. 4. Transient display of spatial frequency information of grating reso-
lution chart captured at times (a) t =0 and (b) t =5 s.
5838 Appl. Phys. Lett., Vol. 85, No. 24, 13 December 2004 Kothapalli et al.
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